Antifungal compounds

ABSTRACT

The technical field of the invention is in pharmaceutical compounds and methods. In an aspect, the disclosure provides macrolide compounds suitable for use as antifungal agents, as well as methods for their use and compositions containing the same.

BACKGROUND

Tacrolimus (also referred to as FK-506) is a compound known to haveimmunosuppressive activity. As an immunosuppressive, it is used in avariety of situations such as organ transplantations and eczematreatment. The structure of tacrolimus includes a macrocyclic lactone,and various structurally related macrolide compounds are known.

Relevant art: US 2006/0035918; U.S. Pat. No. 5,457,111.

SUMMARY OF THE INVENTION

In an aspect is a method for treating a patient suffering from a fungalinfection, the method comprising administering to the patient aneffective amount of a composition comprising a compound of formula (I)

In formula (I), “a” is a double bond optionally present (provided thatR⁵ or R^(5a) is not present); R¹ is selected from alkyl, alkenyl, or istaken together with R³ or R^(3a) to form a cycle; R³ and R^(3a) areindependently selected from —H and —OH, or R³ and R^(3a) together form═X, where X is selected from O, C, and N such that ═X and the carbonatom to which it is attached forms a carbonyl, oxime, substituted oxime,imine, substituted imine, hydrazone, substituted hydrazone, terminalolefin, or substituted olefin functional group, or wherein one of R³ andR^(3a) is taken together with R¹ or R⁵ or R^(5a) to form a cycle (andthe other is H); R⁵ and R^(5a) are independently selected from —H, —OH,or —OTBS, or R⁵ and R^(5a) together form ═O, or one of R⁵ and R^(5a) is—H and the other is taken together with R³ or R^(3a) to form a cycle; R⁷and R^(7a) are independently selected from —H, —OH, —NH₂, alkoxy,alkylcarboxy, alkenylcarboxy, and substituted versions thereof, or R⁷and R^(7a) together form ═Y, where Y is selected from O, and N such that═Y and the carbon atom to which it is attached forms a carbonyl or oximefunctional group; and R9 is selected from —H and —OH.

In embodiments:

the compound has the structure of formula (IA-a), (IA-b), or (IA-c)

the compound has the structure of formula (IB-a), (IB-b), (IB-c), or(IB-d)

wherein ═X is selected from ═C(R^(3b))(R^(3c)), ═N—OR^(3d),═N—NH(R^(3e)), and ═N—N═C(CH₃)₂, R^(3b) and R^(3c) are independentlyselected from —H, —CN, and unsubstituted alkyl, R^(3d) is selected fromH, alkyl, aralkyl, and a function group, and R^(3e) is alkyl;

the compound has the structure of formula (IC-a), (IC-b), (IC-c), or(IC-d)

the compound has the structure of formula (ID) or (IE)

and

the compound has the structure of formula (IF), (IG), or (IH)

In an aspect is an anti-fungal formulation comprising an effectiveamount of a compound having the structure of formula (I) as describedabove, and further comprising a second antifungal agent.

In an embodiment, the second antifungal agent is selected from compoundsaccording to formula (I), polyenes, imidazoles, triazoles, thiazoles,allylamines, and echinochandins.

In an embodiment, there is provided a compound according to any of thestructures described herein.

These and other aspects of the invention will be apparent to the skilledartisan based on the disclosure herein. The technical field of theinvention is in pharmaceutical compounds and methods.

DETAILED DESCRIPTION OF PARTICULAR EMBODIMENTS

The term “alkyl” as used herein refers to a branched, unbranched orcyclic saturated hydrocarbon group of 1 to about 50 carbon atoms, suchas methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl,octyl, decyl, tetradecyl, hexadecyl, eicosyl, tetracosyl and the like.Preferred alkyl groups herein may contain 1 to about 36, more typically1 to 10, carbon atoms. The alkyl groups described herein may beunsubstituted or they may be substituted with one or more substituentsincluding functional groups (e.g., amine, hydroxyl, an olefinic groupsuch as a vinyl or an allyl group), or the like. “Substituted alkyl”refers to alkyl substituted with one or more substituent groups, andthis includes instances wherein two hydrogen atoms from the same carbonatom in an alkyl are replaced, such as in a carbonyl group (i.e., asubstituted alkyl group may include a —C(═O)— moiety). Othersubstituents include halogen, ether, hydroxyl, amine functional groups,etc. as defined in more detail below (see “functional groups”). Theterms “heteroatom-containing alkyl” and “heteroalkyl” refer to an alkylsubstituent in which at least one carbon atom is replaced with aheteroatom, such as O, S, P, or N, as described in further detail infra.If not otherwise indicated, the term “alkyl” includes linear, branched,cyclic, unsubstituted, substituted, heteroatom-containing, andsubstituted heteroatom-containing alkyl.

The term “alkylene” as used herein refers to a difunctional saturatedbranched or unbranched hydrocarbon chain containing from 1 to 50 carbonatoms, more typically from 1 to 12 carbon atoms, and includes, forexample, methylene (—CH₂—), ethylene (—CH₂CH₂—), propylene(—CH₂CH₂CH₂—), 2-methylpropylene (—CH₂—CH(CH₃)—CH₂—), hexylene(—(CH₂)₆-) and the like. Similarly, the terms “alkenylene,”“alkynylene,” “arylene,” “alkarylene,” and “aralkylene” refer todifunctional (i.e., linking) alkenyl, alkynyl, aryl, alkaryl, andaralkyl groups, respectively.

The term “alkenyl” as used herein refers to a linear, branched or cyclichydrocarbon group of 2 to about 50 carbon atoms containing at least onedouble bond, such as ethenyl, n-propenyl, isopropenyl, n-butenyl,isobutenyl, octenyl, decenyl, tetradecenyl, hexadecenyl, eicosenyl,tetracosenyl, and the like. Generally, although again not necessarily,alkenyl groups herein may contain 2 to about 36 carbon atoms, and forexample may contain 2 to 12 carbon atoms, or more typically 2 to 6carbon atoms. The term “substituted alkenyl” refers to alkenylsubstituted with one or more substituent groups, and the terms“heteroatom-containing alkenyl” and “heteroalkenyl” refer to alkenyl inwhich at least one carbon atom is replaced with a heteroatom. If nototherwise indicated, the term “alkenyl” includes linear, branched,cyclic, unsubstituted, substituted, heteroatom-containing, andsubstituted heteroatom containing alkenyl.

The term “alkynyl” as used herein refers to a linear or branchedhydrocarbon group of 2 to 50 carbon atoms containing at least one triplebond, such as ethynyl, n-propynyl, and the like. Generally, althoughagain not necessarily, alkynyl groups herein may contain 2 to about 18carbon atoms, and such groups may further contain 2 to 12 carbon atoms,or more typically 2 to 6 carbon atoms. The term “substituted alkynyl”refers to alkynyl substituted with one or more substituent groups, andthe terms “heteroatom-containing alkynyl” and “heteroalkynyl” refer toalkynyl in which at least one carbon atom is replaced with a heteroatom.If not otherwise indicated, the term “alkynyl” includes linear,branched, unsubstituted, substituted, and/or heteroatom-containingalkynyl.

The term “aryl” as used herein refers to an aromatic species having 1 to3 rings, but typically intends a monocyclic or bicyclic moiety, e.g.,phenyl or 1- or 2-naphthyl groups. Optionally, these groups aresubstituted with 1 to 4, more preferably 1 to 2, substituents such asthose described herein, including alkyl, alkoxy, hydroxyl, amino, and/ornitro. Aryl groups may, for example, contain 6 to 50 carbon atoms, andas a further example, aryl groups may contain 6 to 12 carbon atoms. Forexample, aryl groups may contain one aromatic ring or two fused orlinked aromatic rings, e.g., phenyl, naphthyl, biphenyl, diphenylether,diphenylamine, benzophenone, and the like. “Substituted aryl” refers toan aryl moiety substituted with one or more substituent groups, and theterms “heteroatom-containing aryl” and “heteroaryl” refer to arylsubstituent, in which at least one carbon atom is replaced with aheteroatom. If not otherwise indicated, the term “aryl” includesunsubstituted, substituted, heteroatom-containing, and substitutedheteroatom-containing aromatic substituents.

The term “aralkyl” refers to an alkyl group with an aryl substituent,and the term “alkaryl” refers to an aryl group with an alkylsubstituent, wherein “alkyl” and “aryl” are as defined above. Ingeneral, aralkyl and alkaryl groups herein contain 6 to 50 carbon atoms.Aralkyl and alkaryl groups may, for example, contain 6 to 20 carbonatoms, and as a further example, such groups may contain 6 to 12 carbonatoms. Unless specified otherwise, the terms “alkaryl” and “aralkyl”include substituted, heteroatom-containing, and substitutedheteroatom-containing versions thereof.

The term “amino” intends an amino group —NR₂ where R is hydrogen or analternative substituent, typically alkyl. The term “amino” is thusintended to include primary amino (i.e., NH₂), “alkylamino” (i.e., asecondary amino group containing a single alkyl substituent), and“dialkylamino” (i.e., tertiary amino group containing two alkylsubstituents).

The term “heteroatom-containing” as in a “heteroatom-containing alkylgroup” (also termed a “heteroalkyl” group) or a “heteroatom-containingaryl group” (also termed a “heteroaryl” group) refers to a molecule,linkage or substituent in which one or more carbon atoms are replacedwith an atom other than carbon, e.g., nitrogen, oxygen, sulfur,phosphorus or silicon, typically nitrogen, oxygen or sulfur. Similarly,the term “heteroalkyl” refers to an alkyl substituent that isheteroatom-containing, the term “heterocyclic” refers to a cyclicsubstituent that is heteroatom-containing, the terms “heteroaryl” andheteroaromatic” respectively refer to “aryl” and “aromatic” substituentsthat are heteroatom-containing, and the like. Examples of heteroalkylgroups include alkoxyaryl, alkylsulfanyl-substituted alkyl, N-alkylatedamino alkyl, and the like. Examples of heteroaryl substituents includepyrrolyl, pyrrolidinyl, pyridinyl, quinolinyl, indolyl, furyl,pyrimidinyl, imidazolyl, 1,2,4-triazolyl, tetrazolyl, etc., and examplesof heteroatom-containing alicyclic groups are pyrrolidino, morpholino,piperazino, piperidino, tetrahydrofuranyl, etc.

“Halo” or “halogen” refers to fluoro, chloro, bromo or iodo, and usuallyrelates to halo substitution for a hydrogen atom in an organic compound.

By “substituted” as in “substituted hydrocarbyl,” “substituted alkyl,”“substituted aryl,” and the like, as alluded to in some of theaforementioned definitions, is meant that in the hydrocarbyl, alkyl,aryl, or other moiety, at least one hydrogen atom bound to a carbon (orother) atom is replaced with one or more non-hydrogen substituents.Examples of such substituents include, without limitation: C₁-C₂₄ alkyl(including C₁-C₁₈ alkyl, further including C₁-C₁₂ alkyl, and furtherincluding C₁-C₆ alkyl), C₂-C₂₄ alkenyl (including C₂-C₁₈ alkenyl,further including C₂-C₁₂ alkenyl, and further including C₂-C₆ alkenyl),C₂-C₂₄ alkynyl (including C₂-C₁₈ alkynyl, further including C₂-C₁₂alkynyl, and further including C₂-C₆ alkynyl), C₅-C₃₀ aryl (includingC₅-C₂₀ aryl, and further including C₅-C₁₂ aryl), C₆-C₃₀ aralkyl(including C₆-C₂₀ aralkyl, and further including C₆-C₁₂ aralkyl), C₆-C₃₀alkaryl (including C₆-C₂₀ alkaryl, and further including C₆-C₁₂alkaryl), and functional groups such as halo, hydroxyl, sulfhydryl,C₁-C₂₄ alkoxy, C₂-C₂₄ alkenyloxy, C₂-C₂₄ alkynyloxy, C₅-C₂₀ aryloxy,acyl (including C₂-C₂₄ alkylcarbonyl (—CO-alkyl) and C₆-C₂₀ arylcarbonyl(—CO-aryl)), acyloxy (—O-acyl), C₂-C₂₄ alkoxycarbonyl (—(CO)—O-alkyl),C₆-C₂₀ aryloxycarbonyl (—(CO)—O-aryl), halocarbonyl (—CO)—X where X ishalo), C₂-C₂₄ alkylcarbonato (—O—(CO)—O-alkyl), C₆-C₂₀ arylcarbonato(—O—(CO)—O-aryl), C₂-C₂₄ alkylcarbonyloxy (—O—(CO)-alkyl), C₆-C₂₄arylcarbonyloxy (—O—(CO)-aryl), carboxy (—COOH), carboxylato (—COO—),carbamoyl (—(CO)—NH₂), mono-substituted C₁-C₂₄ alkylcarbamoyl(—(CO)—NH(C₁-C₂₄ alkyl)), di-substituted alkylcarbamoyl (—(CO)—N(C₁-C₂₄alkyl)₂), mono-substituted arylcarbamoyl (—(CO)—NH-aryl), thiocarbamoyl(—(CS)—NH₂), carbamido (—NH—(CO)—NH₂), cyano (—C≡N), isocyano (—N+≡C—),cyanato (—O—C≡N), isocyanato (—O—N≡C—), isothiocyanato (—S—C≡N), azido(—N═N+=N—), formyl (—(CO)—H), thioformyl (—(CS)—H), amino (—NH₂), mono-and di-(C₁-C₂₄ alkyl)-substituted amino, mono- and di-(C₅-C₂₀aryl)-substituted amino, C₂-C₂₄ alkylamido (—NH—(CO)-alkyl), C₅-C₂₀arylamido (—NH—(CO)-aryl), imino (—CR═NH where R=hydrogen, C₁-C₂₄ alkyl,C₅-C₂₀ aryl, C₆-C₂₀ alkaryl, C₆-C₂₀ aralkyl, etc.), alkylimino(—CR═N(alkyl), where R=hydrogen, alkyl, aryl, alkaryl, etc.), arylimino(—CR═N(aryl), where R=hydrogen, alkyl, aryl, alkaryl, etc.), nitro(—NO₂), nitroso (—NO), sulfo (—SO₂—OH), sulfonato (—SO₂—O—), C₁-C₂₄alkylsulfanyl (—S-alkyl; also termed “alkylthio”), arylsulfanyl(—S-aryl; also termed “arylthio”), C₁-C₂₄ alkylsulfinyl (—(SO)-alkyl),C₅-C₂₀ arylsulfinyl (—(SO)-aryl), C₁-C₂₄ alkylsulfonyl (—SO₂-alkyl),C₅-C₂₀ arylsulfonyl (—SO₂-aryl), phosphono (—P(O)(OH)₂), phosphonato(—P(O)(O—)₂), phosphinato (—P(O)(O—)), phospho (—PO₂), phosphino (—PH₂),mono- and di-(C₁-C₂₄ alkyl)-substituted phosphino, and mono- anddi-(C₅-C₂₀ aryl)-substituted phosphino. In addition, the aforementionedfunctional groups may, if a particular group permits, be furthersubstituted with one or more additional functional groups or with one ormore hydrocarbon moieties (alkyl, aryl, etc.). Analogously, theabove-mentioned hydrocarbon moieties may be further substituted with oneor more functional groups or additional hydrocarbon moieties such asthose specifically enumerated. It will be appreciated that functionalgroups may be attached via a heteroatom or, where appropriate, via acarbon atom, to the remainder of the compound.

In an aspect is a compound having the structure of formula (I)

In formula (I), “a” is a double bond optionally present (provided thatR⁵ or R^(5a) is not present); R¹ is selected from alkyl, alkenyl, or istaken together with R³ or R^(3a) to form a cycle; R³ and R^(3a) areindependently selected from —H and —OH, or R³ and R^(3a) together form═X, where X is selected from O, C, and N such that ═X and the carbonatom to which it is attached forms a carbonyl, oxime, substituted oxime,imine, substituted imine, hydrazone, substituted hydrazone, terminalolefin, or substituted olefin functional group, or wherein one of R³ andR^(3a) is taken together with R¹ or R⁵ or R^(5a) to form a cycle (andthe other is H); R⁵ and R^(5a) are independently selected from —H, —OH,or —OTBS, or R⁵ and R^(5a) together form ═O, or one of R⁵ and R^(5a) is—H and the other is taken together with R³ or R^(3a) to form a cycle; R⁷and R^(7a) are independently selected from —H, —OH, —NH₂, alkoxy,alkylcarboxy, alkenylcarboxy, and substituted versions thereof, or R⁷and R^(7a) together form ═Y, where Y is selected from O, and N such that═Y and the carbon atom to which it is attached forms a carbonyl or oximefunctional group; and R⁹ is selected from —H and —OH.

In formula (I), R¹ is selected from alkyl and alkenyl, or R¹ may betaken together with R³ or R^(3a) to form a cycle. Examples of alkylgroups include methyl and substituted methyl (e.g., —C(═O)-Me,—C(═O)—OH, and —C(═O)—OMe), while examples of alkenyl groups include—CH═CR^(1c)R^(1d). Where R¹ is —CH═CR^(1c)R^(1d), the double bond may bein the E- or Z-configuration, and the formulation may comprise a singleisomer or a mixture of isomers. In embodiments, R¹ is unsubstitutedalkyl or unsubstituted alkenyl.

R^(1c) and R^(1d) are independently selected from: H, alkyl, aryl,alkaryl, aralkyl, and a functional group. Examples include —(CH₂)_(n)CH₃where n is an integer (e.g., an integer in the range 0-20, or an integerselected from 0, 1, 2, 3, 4, 5, etc.), cyclohexyl, substituted alkyl(substituents such as aryl and functional groups), phenyl, substitutedphenyl (substituents such as alkyl, alkenyl, functional groups, etc.),alkoxycarbonyl (e.g., C(═O)O-alkyl and C(═O)O-aryl), alkylsulfonyl(e.g., —SO₂-Me or —SO₂-Et), etc.

In embodiments, R¹ is alkyl including branched alkyl, such as methyl,ethyl, i-propyl, i-butyl, t-butyl, etc.

In any of the embodiments of formula (I) described herein where R¹ isnot part of a cycle, R¹ may be selected from -Me and —CH═CH₂.

In embodiments, R¹ is taken together with R³ or R^(3a) to formsubstituted or unsubstituted pyridazine.

In embodiments of formula (I), R³ and R^(3a) are independently selectedfrom —H or —OH, or R³ and R^(3a) are taken together to form ═X, where Xis O, N, or C such that X and the carbon atom to which it is attachedform carbonyl, oxime, substituted oxime, imine, substituted imine,hydrazone, or substituted hydrazone, or X is C to form an olefin(terminal or internal). In embodiments, ═X is selected from ═O,═C(R^(3b))(R^(3c)), ═N—OR^(3d), ═N—N═C(CH₃)₂, and ═N—NH—R^(3e), orwherein R³ or R^(3a) is taken together with R¹ or R⁵ or R^(5a) to form acycle. In embodiments, R³ and R^(3a) together are ═O;═C(R^(3b))(R^(3c)); or ═N—OR^(3d). In embodiments, R³ and R^(3a)together are ═O or ═C(R^(3b))(R^(3C)). In embodiments, R³ and R^(3a)together are ═N—OR^(3d), with two isomers present for the possibleorientations of the —OR^(3d) group. The compound may be racemic (withboth isomers) or may be a single oxime isomer in the formulationsdescribed herein.

In embodiments, R³ and R^(3a) are both H.

In embodiments, R^(3b) and R^(3c) are independently selected from H andunsubstituted alkyl. In embodiments R^(3b) and R^(3c) are both H. Inembodiments exactly one of R^(3b) and R^(3c) is H and the other isunsubstituted alkyl. In embodiments both R^(3b) and R^(3c) areunsubstituted alkyl. Examples of alkyl include methyl, ethyl, propyl(n-propyl or i-propyl), butyl (n-butyl, i-butyl, t-butyl), pentyl, andhexyl. In other embodiments, one of R^(3b) and R^(3c) is H, and theother is —CN.

R^(3d) is selected from H, alkyl, aralkyl, and a function group.Examples of alkyl include methyl, ethyl, propyl (i.e., n- and i-propyl),butyl (i.e., n-, i-, and t-butyl), —(CH₂)_(n)—CH₃ (wherein n is in therange 1-5 or 1-3, or is 1, 2, 3, 4, or 5), —CH₂—COOH, and —(CH₂)_(n)—OH(wherein n is in the range 1-5 or 2-4, or is 1, 2, 3, 4, or 5). Examplesof aralkyl include —CH₂—C₆H₄—NO₂ and —CH₂—C₆H₃Cl₂. Examples offunctional groups include —S(═O)₂—OH.

R^(3e) is alkyl. Examples of alkyl include methyl, ethyl, propyl (i.e.,n- and i-propyl), butyl (i.e., n-, i-, and t-butyl), —(CH₂)_(n)—CH₃(wherein n is in the range 1-5 or 1-3, or is 1, 2, 3, 4, or 5),—(CH₂)_(n)—OH (wherein n is in the range 1-5 or 2-4, or is 1, 2, 3, 4,or 5), etc.

In formula (I), in embodiments, bond “a” is present and R^(5a) is notpresent. In other embodiments, bond “a” is not present and R^(5a) ispresent. In some such embodiments, R^(5a) is H.

In formula (I), one of R⁵ and R^(5a) is —OH or —OTBS and the other is H,or R⁵ and R^(5a) taken together form ═O, or R⁵ is taken together with R³to form a cycle. In embodiments, R³ or R^(3a) and R⁵ or R^(5a) are takentogether to form a cycle such as a ketal or acetal (e.g., adimethylacetonide).

In formula (I), one of R⁷ and R^(7a) is selected from —OH, —NH₂,alkylcarboxy (e.g., —O—CO—CH₂—COOH), alkenylcarboxy (e.g.,—O—CO—CH₂CH₂CH═CH₂, —O—CO—CH₂CH₂CH═CH—COOH etc.), and thiocarbonato(e.g., —O—C(═S)—O—R where R is alkyl or aryl). Alternatively, R⁷ andR^(7a) together form ═Y, where ═Y is N or O to form an oxime or carbonylgroup.

In embodiments, R⁷ and R^(7a) are both —H.

In embodiments, the compounds have the structure of formula (IA-a),(IA-b), or (IA-c)

In formula (IA-a), (IA-b), and (IA-c), one of R³ and R^(3a) is H and theother is taken with R¹ (formula IA-c) or with R⁵ or R^(5a) to form acycle; and R⁷, and R^(7a) are as defined for formula (I).

For example, in formula (IA-a) and (IA-b), R⁷ and R^(7a) are OH and H,respectively, R³ and R^(5a) are —H, and R^(3a) and R⁵ are taken togetherto form a cycle. An example cycle is an acetonide group. Alternatively,R³ and R⁵ are H, and R^(3a) and R^(5a) are taken together to form anacetonide or other cycle. Alternatively, R^(3a) and R⁵ are H, and R³ andR^(5a) are taken together to form an acetonide or other cycle.Alternatively, R^(3a) and R^(5a) are H, and R³ and R⁵ are taken togetherto form an acetonide or benzylidene acetal (i.e. —O—C(H)(Ph)-O— wherethe oxygen atoms are connected at C22 and C24). In such compounds, R⁷and R^(7a) may alternatively both be —H, or may together form ═O.

For example, in formula (IA-c), R⁷ and R^(7a) are OH and H,respectively, R⁵ and R^(5a) are OH and H, respectively, and R³ or R^(3a)is taken together with R¹ to form a substituted or unsubstitutedpyridazine cycle (the other of R³ and R^(3a) being H). Examplesubstituents are alkyl.

In embodiments, the compounds have the structure of formula (IB-a) or(IB-b) or (IB-c) or (IB-d):

In embodiments of formula (IB-a), (IB-b), (IB-c), and (IB-d), ═X isselected from ═C(R^(3b))(R^(3c)), ═N—OR^(3d), ═N—NH(R^(3e)), and═N—N═C(CH₃)₂, R^(3b), R^(3c), R^(3d), R^(3e), R⁵, R⁷, and R^(7a) are asdefined previously for formula (I).

For example, in formula (IB-a), (IB-b), (IB-c), and (IB-d), R⁵ is —OH,R⁷ and R^(7a) are OH and H, respectively, and X is ═CH₂.

For example, in formula (IB-a), (IB-b), (IB-c), and (IB-d), R⁵ is —OH,R⁷ and R^(7a) are OH and H, respectively, and X is ═N—OH.

For example, in formula (IB-a), (IB-b), (IB-c), and (IB-d), R⁵ is —OH,R⁷ and R^(7a) are OH and H, respectively, and X is ═N—O—R^(3d). Forexample, R⁵ is —OH, R⁷ and R^(7a) are OH and H, respectively, and X isselected from ═N—O—CH₃, ═N—O—(CH₂)_(n)—CH₃ (n=1, 2, or 3),═N—O—CH(CH₃)₂, ═N—O—C(CH₃)₃, ═N—O—(CH₂)_(n)—OH (n=1, 2, or 3), and═N—O—(CH₂)_(n)—COOH (n is 1, 2, or 3). Or for example, R⁵ is —OH, R⁷ andR^(7a) are OH and H, respectively, and X is ═N—O—CH₂-aryl, where aryl isphenyl, nitrophenyl (e.g., 4-nitrophenyl), or halophenyl (e.g.,chlorophenyl such as 4-chlorophenyl, dichlorophenyl such as2,4-dichlorophenyl, and trichlorophenyl such as 2,4,6-trichlorophenyl).

For example, in formula (IB-a), (IB-b), (IB-c), and (IB-d), R⁵ is —H or—OH, R⁷ and R^(7a) are OH and H, respectively, and X is ═C(H)(CN). Alsofor example, X is ═C(Me)(Et).

For example, in formula (IB-a), (IB-b), (IB-c), and (IB-d), R⁵ is —H or—OH, R⁷ and R^(7a) are OH and H, respectively, X is ═N—NH—R^(3e), andR^(3e) is selected from methyl, ethyl, i-propyl, n-propyl, and—(CH₂)_(n)—OH (n is 0, 1, 2, or 3).

For example, in formula (IB-a), (IB-b), (IB-c), and (IB-d), R⁵ is —H or—OH, R⁷ and R^(7a) are OH and H, respectively, X is ═N—N═C(CH₃)₂.

For example, in formula (IB-a), (IB-b), (IB-c), and (IB-d), R⁵ is —H or—OH, R⁷ and R^(7a) are OH and H, respectively, X is ═N—O—SO₂—H or═N—O—SO₂—R where R is alkyl such as Me.

For example, in formula (IB-a), (IB-b), (IB-c), and (IB-d), R⁵ is —H or—OH, R⁷ and R^(7a) are taken together to form ═N—OH, and X is ═N—OH.

For example, in formula (IB-a), (IB-b), (IB-c), and (IB-d), one of R⁷and R^(7a) is —H and the other is —OH, R⁵ is —H, —OH, or —OTBS, and X is═N—OH, ═N—OR^(3j), or ═N—NHMe, where R^(3j) is alkyl such as methyl,ethyl, propyl, butyl, etc.

In embodiments of formula (IB-a), (IB-b), (IB-c), and (IB-d), R⁷ andR^(7a) are —H, R⁵ is —H or —OH, and X is ═N—OH, ═N—OR^(3j), or ═N—NHMe,where R^(3j) is alkyl such as methyl, ethyl, propyl, butyl, etc.

In embodiments of formula (IB-a), (IB-b), (IB-c), and (IB-d), R⁵ is —Hor —OH, X is ═CH₂ or ═O, and R⁷ and R^(7a) together form ═O.

In embodiments of formula (IB-a), (IB-b), (IB-c), and (IB-d), R⁷ andR^(7a) are —OH and —H, respectively (or, they are both —H), R⁵ is —H or—OH, and X is ═C(R^(3b))(R^(3c)), where R^(3b) and R^(3c) areindependently selected from —H, —CN, and alkyl.

In each of the above oxime and alkene compounds for formula (IB-a),(IB-b), (IB-c), and (IB-d), the oxime/alkene may exist as a racemicmixture of two isomers, or may be present as a single isomer. Isolationof single isomers is generally within the skill in the art.

In embodiments of formula (IB-a), (IB-b), (IB-c), and (IB-d), R⁵ is —OHor —H, X is ═O and R⁷ and R^(7a) are both H.

In any of the foregoing embodiments of formula (IB-a), (IB-b), (IB-c),and (IB-d), R⁷ and R^(7a) may alternatively together form ═O, or mayalternatively both be —H.

In embodiments, the compounds have the structure of formula (IC-a),(IC-b), (IC-c), or (IC-d)

In Formula (IC-a), (IC-b), (IC-c), and (IC-d), R³, R^(3a), R⁷, andR^(7a) are as defined in Formula (I). In embodiments, R³ and R^(3a)together form ═O, one of R⁷ and R^(7a) is —H, and the other is selectedfrom —OH, alkoxy, alkylcarboxy, alkenylcarboxy, and substituted versionsthereof.

For example, in formula (IC-a), (IC-b), (IC-c), and (IC-d), R³ andR^(3a) together form ═O, R^(7a) is —H, and R⁷ is —OH or —O—C(═O)—R^(7c),where R^(7c) is —(CH₂)_(n)—COOH or —(CH₂)_(n)—CH═CHR^(7d) (n is 1, 2, or3), and R^(7d) is —H, alkyl (e.g., methyl, ethyl, etc), or —COOH.

For example, in formula (IC-a), (IC-b), (IC-c), and (IC-d), R^(7a) is—H, R⁷ is —OH and R³ and R^(3a) together form ═N—NH—R^(7e), ═N—OH, or═N—OR^(7e), where R^(7e) is alkyl (e.g., methyl or ethyl, etc.).

In embodiments, the compounds have the structure of formula (ID) or (IE)

In formula (ID) and (IE), R¹, R⁷, R^(7a), and R⁹ are as defined informula (I). The wavy line indicates that the hydroxyl substituent canbe either isomer, and both isomers are intended to be included.

In embodiments of formula (ID) and (IE), R¹ is alkyl or alkenyl, R^(7a)and R⁹ are —H, and R⁷ is selected from alkoxy, alkylcarboxy,alkenylcarboxy, and substituted versions thereof. For example, R¹ ismethyl, R^(7a) and R⁹ are —H, and R⁷ is —OC(═O)—(CH₂)_(n)—COOH (n is 1,2, or 3). For example, R¹ is —CH═CH₂, R^(7a) and R⁹ are —H, and R⁷ is—OC(═O)—(CH₂)_(n)—COOH (n is 1, 2, or 3). For example, R¹ is methyl,R^(7a) and R⁹ are —H, and R⁷ is —OH. For example, R¹ is —CH═CH₂, R^(7a)and R⁹ are —H, and R⁷ is —OH.

In embodiments of formula (ID) and (IE), R¹ is alkyl or alkenyl, R^(7a)is —H, and R⁷ and R⁹ are both hydroxyl. For example, R¹ is methyl. Forexample, R¹ is —CH═CH₂.

In embodiments the compounds have the structure of formula (IF-a) or(IF-b)

In formula (IF), R¹, R⁵, R⁷, and R^(7a) are as defined in formula (I).

In embodiments of formula (IF), R¹ is alkyl or alkenyl, R⁵ is —OH, R⁷ isH, and R^(7a) is amine. For example, R¹ is methyl, R⁵ is —OH, R⁷ is H,and R^(7a) is —NH₂. For example, R¹ is —CH═CH₂, R⁵ is —OH, R⁷ is H, andR^(7a) is —NH₂. For example, R¹ is methyl or —CH═CH₂, R⁵ is —OH, R^(7a)is H, and R⁷ is —NH₂.

In embodiments of formula (IF), R¹ is alkyl or alkenyl, R⁵ is —OTBS, R⁷is H, and R^(7a) is amine. For example, R¹ is methyl, R⁵ is —OTBS, R⁷ isH, and R^(7a) is —NH₂. For example, R¹ is —CH═CH₂, R⁵ is —OTBS, R⁷ is H,and R^(7a) is —NH₂. For example, R¹ is methyl or —CH═CH₂, R⁵ is —OTBS,R^(7a) is H, and R⁷ is —NH₂. Also for example, R¹ is methyl or —CH═CH₂,R⁵ is —OTBS, R^(7a) is H, and R⁷ is —H.

In embodiments of formula (IF), R¹ is alkyl or alkenyl, R⁵ is —OH, andR⁷ and R^(7a) are —H. For example, R¹ is methyl or —CH═CH₂, R⁵ is —OH,and R⁷ and R^(7a) are —H.

In embodiments of formula (IF), R¹ is alkyl or alkenyl, R⁵ is —OH, andR⁷ and R^(7a) are thiocarbonato and —H, respectively. For example, R¹ ismethyl or —CH═CH₂, R⁵ is —OH, R^(7a) is H, and R⁷ is —O—C(═S)—O-alkyl or—O—C(═S)—O-aryl. For example, R⁷ is —O—C(═S)—O-methyl or —O—C(═S)—O-Ph.

In embodiments of formula (IF), R⁵ is —OH, and R⁷ and R^(7a) togetherform ═O, and R¹ is selected from alkenyl. For example, R⁵ is —OH, and R⁷and R^(7a) together form ═O, and R¹ is —CH═CH—(CH₂)_(n)—R^(7f), where nis 1, 2, 3, 4, 5, or greater than 5, and R^(7f) is methyl, —OH, alkoxy,or aryloxy. For example, R⁵ is —OH, and R⁷ and R^(7a) together form ═O,and R¹ is selected from —CH═CH—(CH₂)_(n)—O—R^(7g), where n is 1, 2, 3,or 4, and R^(7g) is methyl, —OH, —OMe, or —OPh.

In embodiments of formula (IF), R⁵ is —OH, and R⁷ and R^(7a) are —OH and—H, respectively, and R¹ is selected from alkyl and alkenyl. Forexample, R¹ is methyl or —CH═CH₂.

In embodiments, the compounds have the structure of formula (IG)

In formula (IG), R¹, R⁷, and R^(7a) are as defined in formula (I).

For example, R¹ is selected from alkyl and alkenyl, one of R⁷ and R^(7a)is —H, and the other is selected from —H and —OH. For example, R⁷ is—OH, R^(7a) is —H, and R¹ is methyl, or —CH═CH₂. Also for example, R⁷and R^(7a) together form ═O, and R¹ is selected from alkyl and alkenyl(e.g., -Me, —CH═CH₂, etc.). Also for example, both R⁷ an R^(7a) are —H.

In embodiments, the compounds have the structure of formula (IH)

In formula (IH), R¹ is as defined in formula (I).

For example, R¹ is alkyl, including substituted methyl. For example, R¹is methyl, —C(═O)-Me, —C(═O)—OH or —CHR^(1a)R^(1b), wherein one ofR^(1a) and R^(1b) is —H and the other is selected from —H, carboxylicacid, and alkylcarbonyl such as —C(═O)Me or —C(═O)Et.

For example, R1 is alkenyl, including substituted alkenyl. Examplesinclude —CH═CH₂, —CH═CH(CH₃) (E and Z configuration), —CH═C(CH₃)₂, and—CH═CH(R^(1e)) wherein R^(1e) is alkyl. Examples of R^(1e) include—(CH₂)_(n)—R^(1f) (where n is in the range 1-20, or 1-10, and R^(if) isMe or —OH), acetals, alkyl groups substituted with sufone andsulfonyloxy groups, alkyl groups substituted with ester or carbonyloxygroups, cyclic alkyl groups including heterocyclic alkyl groups, arylgroups including heterocyclic aryl groups, heteroatoms substituted withalkyl groups, ketone groups, amide groups, bicyclic groups includingbicyclic aromatic and bicyclic heteroatom-containing groups, and thelike.

Unless otherwise specified, reference to “formula (I)” includes allsub-formulae of formula (I) (i.e., IA-a, IA-b, IA-c, IB-a, IB-b, etc.).

Included are salts (e.g., pharmaceutically acceptable salts) of thecompounds of formula (I). Examples of salts are halo salts (e.g.,chloride, fluoride, bromide, or iodide salts), fluorinated salts such asperfluoroacetic acid (CF₃COOH) salt, acetic acid salt, and the like.

Examples of specific compounds according to formula (I) are given inTable 1.

In an aspect, the compounds are useful in treating a fungal infection ina patient. Patients include human patients as well as non-human patients(e.g., domesticated animals and the like).

In an aspect, a patient suffering from a fungal infection is treatedwith a formulation containing at least one compound according to aformula herein.

Examples of fungal infections suitable for treatment by formulationsdescribed herein include Candida, Aspergillus, Microsporum,Trichophyton, Cryptococcus, and Epidermophyton.

The compounds disclosed herein may be used as a pharmaceutically activecompound to prepare a pharmaceutically active formulation. Suchformulation may further comprise additives such as pharmaceuticallyacceptable carriers, colorants, flavorants, binders, etc., and mayfurther comprise coatings (if in solid dosage form), solvents (if inliquid oral, spray, or injectable form), and the like.

The total daily dose of the described compounds administered to apatient may range from about 0.001 to about 3 mg/kg/day. For purposes oforal administration, more preferable doses may be in the range of fromabout 0.005 to about 1.5 mg/kg/day. If desired, the effective daily dosemay be divided into multiple doses for purposes of administration;consequently, single dose compositions may contain such amounts orsubmultiples thereof to make up the daily dose.

In embodiments, the formulation comprises a second antifungal agent. Thesecond antifungal agent may be another compound according to theformulae herein. In embodiments, the second antifungal agent is a knownantifungal and not a compound according to the formulae herein, such asa polyene, imidazole, triazole, thiazole, allylamine, echinocandin,among others. Examples include Amphotericin B, Candicidin, Filipin,Hamycin, Natamycin, Nystatin, Rimocidin, Bifonazole, Butoconazole,Clotrimazole, econazole, fenticonazole, isoconazole, kentoconazole,miconazole, omoconazole, oxiconazole, sertaconazole, sulconazole,tioconazole, albaconazole, fluconazole, isavuconazole, itraconazole,posaconazole, ravuconazole, terconazole, voriconazole, abafungin,amorolfin, butenafine, naftifine, terbinafine, anidulafungin,caspofungin, micafungin, benzoic acid, ciclopirox, flucytosine,griseofulvin, haloprogin, polygodial, tolnaftate, undecylenic acid, andcrystal violet, among others.

TABLE 1 Antifungal Compounds

2

3

4

5

6

7

8

9

10

11

12

13

14

15

16

17

18

19

20

21

22

23

24

25

26

27

28

29

30

31

32

33

34

35

36

37

38

39

40

41

42

43

44

45

46

47

48

49

50

51

52

53

54

55

56

57

58

59

60

61

62

63

64

65

66

67

68

69

70

71

72

73

74

75

76

77

78

79

80

81

82

83

84

85

86

87

88

89

90

91

92

93

94

95

96

97

98

99

100

101

102

103

104

105

106

107

108

109

110

111

112

113

114

115

116

117

118

119

120

121

122

123

124

125

126

127

128

129

130

131

132

133

134

135

136

137

138

139

140

141

142

143

144

145

146

147

148

149

150

151

152

153

154

155

156

157

158 159

160

161

162

163

164

165

166

167

168

169

170

171

172

173

174

175

176

177

178

179

180

181

182

183

184

185

186

187

188

189

190

191

192

193

194

195

196

197

198

199

200

201

202

203

204

205

206

207

208

209

210

211

212

213

214

215 216

217

218

219

EXAMPLES

Preparation of “C22”-Oximes from FK506 or Ascomycin.

Example Procedure: Combined FK506 (0.50 g, 0.60 mmol), hydroxylaminehydrochloride (0.50 g, 7.0 mmol), pyridine (0.25 mL, 3.2 mmol), andethanol (60 mL). The mixture was heated at reflux. LCMS at 2 h indicatedcomplete reaction. The mixture was cooled to rt, diluted with water, andtreated with dilute HCl (to ˜pH4). The Ethanol was evaporated, and theresidue was extracted into DCM three times. The combined organic phasewas washed with brine, dried over Na₂SO₄, and evaporated to give a whitesolid. Purification by Biotage flash chromatography (25 g SNAP column,7-60% Acetone/Hexane). Fraction 16 (82 mg, 16%) appears to be enrichedin one oxime isomer, fraction 18 (68 mg, 13%) appears to be enriched inthe other oxime isomer, and fraction 17 (103 mg, 20%) is a less-enrichedmixture of isomers.

Preparation of “C22”-Hydrazones from FK506 or Ascomycin.

Example Procedure: Combined FK506 (0.50 g, 0.62 mmol), Ethanol (35 mL),2-hydroxyethylhydrazine (0.25 mL, 3.7 mmol), and TsOH (0.71 g, 3.7mmol). The mixture was stirred at rt for 20 h. The solvent wasevaporated and the residue was purified by Biotage flash chromatography(25 g SNAP, 7-60% Acetone/Hexane). The appropriate fractions werecombined and further purified by NP-HPLC (Kromasil, 4.6 mm×250 mm, 100-5sil, 20% EtOH/Heptane). The appropriate fractions were combined andevaporated to give the desired material as a white solid (32 mg 6%).

Preparation of C₂₃-C₂₄-dehydro-C22-ethylhydrazone from FK506.

Procedure: Combined FK506 (300 mg, 0.37 mmol), ethanol (21 mL),ethylhydrazine hydrochloride (216 mg, 2.2 mmol), and TsOH (426 mg, 2.2mmol), and the mixture was stirred at rt. LCMS at 18 h indicated nostarting material remained. The mixture was diluted with DCM and waterand then adjusted to neutral pH with NaHCO₃. The organic solvents wereevaporated and the aqueous residue was extracted with DCM three times.The combined organics were washed with brine, dried over Na₂SO₄, andevaporated to give an oil. Purification with Biotage flashchromatography (25 g SNAP, 7-60% acetone/hexanes). Both theC24-hydroxy-C22-hydrazone (24 mg, 8%) and theC₂₃-C₂₄-dehydro-C22-hydrazone (22 mg, 7%) were isolated from a separablemixture.

Preparation of C22 Exocyclic Alkenes from Ascomycin.

Example Procedure using the Peterson Olefination:C24,C32-bis-TBS-protected ascomycin (0.18 g, 0.18 mmol) was dissolved inTHF (5 mL) and the solution was cooled to −78° C. TMSCH₂Li (0.44 mL of1M solution in pentane, 0.44 mmol) was added dropwise (a yellow colorappeared and dissipated with each drop, and then an orange color finallypersisted. The mixture was maintained at −78° C. for 20 h. The reactionwas quenched at -78° C. with two drops of glacial acetic acid. Water wasadded and the mixture was brought to rt. The mixture was treated with asaturated solution of NaHCO₃, and then the pH 8 mixture was extractedwith ether. The ethereal extract was washed with brine, dried overNa₂SO₄, and then evaporated to give an oil. This material was thendissolved in acetonitrile (4.5 mL), and treated with a 48% aqueous HFsolution (0.50 mL, 14 mmol). The mixture was stirred for 8 h, and thenquenched by the addition of ethoxytrimethylsilane (2.0 mL, 12.8 mmol).The mixture was evaporated to dryness and purified by Biotage flashchromatography (10 g SNAP column, 7-60% acetone/hexanes). The productcontaining fractions were combined and further purified by normal phaseHPLC (Kromasil, 4.6 mm×250 mm, 100-5 sil). The appropriate fractionswere combined to give the desired product as a glassy solid (9.2 mg,7%).

Example Procedure using a disubstituted alkyl lithium:C24,C32-bis-TBS-protected ascomycin (0.29 g, 0.28 mmol) was dissolved inTHF (8 mL) and the solution was cooled to −78° C. Sec-butyl lithium(0.50 mL of a 1.4 M solution in cyclohexane, 0.70 mmol) was addeddropwise (a yellow color appeared and dissipated with each drop, andthen an orange color finally persisted. The mixture was maintained at−78° C. for 24 h. The reaction was quenched at −78° C. with 3 drops ofglacial acetic acid. Water was added and the mixture was brought to rt.The mixture was treated with a saturated solution of NaHCO₃, and thenthe pH 8 mixture was extracted with ether. The ethereal extract waswashed with brine, dried over Na₂SO₄, and then evaporated to give anoily white solid. This material was then dissolved in acetonitrile (7.5mL), and treated with a 48% aqueous HF solution (0.80 mL, 22 mmol). Themixture was stirred for 8 h, and then quenched by the addition ofethoxytrimethylsilane (2.0 mL, 12.8 mmol). The mixture was evaporated todryness and purified by Biotage flash chromatography (10 g SNAP column,7-60% acetone/hexanes). The product containing fraction was furtherpurified by normal phase HPLC (Kromasil, 4.6 mm×250 mm, 100-5 sil). Theappropriate fractions were combined to give the desired product (11 mg,5%).

Preparation of C22,C24-Acetonide from Ascomycin.

Procedure: Dissolved Me₄N(OAc)₃BH (0.80 g, 0.30 mmol) in ACN (1 mL) andglacial acetic acid (1.5 mL) at rt. Cooled to 0° C. and stirred for 10min, then added a solution of ascomycin (0.30 g, 0.38 mmol) dissolved inACN (1.5 mL) and EtOAc (1 mL). The vial was sealed and the mixturestirred at 0° C. LCMS after 2 h indicated no starting material remained.The desired m/z was present along with the m/z corresponding toover-reduction. The reaction was quenched with Rochelle's Salt (0.5 M, 2mL) and the mixture was transferred to a round bottom flask andevaporated. The residue was extracted with ethyl acetate (3×25 mL). Thecombined organic phase was washed with brine, dried over sodium sulfate,and then the solvent was evaporated. The crude product was dissolved inacetone (13 mL) and 2,2-dimethoxypropane (13 mL), and then a catalyticamount of pyridinium p-toluenesulfonate was added (10 mg, 0.038 mmol).The mixture was stirred at rt, and after one day there was no remainingstarting material by TLC. The mixture was diluted with water, and thensaturated sodium bicarbonate solution was added (1 mL). The solventswere evaporated and the aqueous residue was extracted with DCM (3×25mL). The combined organic phase was washed with brine and dried oversodium sulfate. The solvent was evaporated to give a white solid whichwas purified by Biotage Isolera flash chromatography (10 g SNAP column,5-40-65% (ethyl acetate/hexanes) step-gradient. The appropriatefractions were combined to give the desired product as a white solid (19mg, 6%).

Preparation of C23-C24-Dehydro-C22-Methyloxime from Ascomycin.

Procedure: Ascomycin (6.8 g, 8.6 mmol) was dissolved in toluene (140 mL)and then p-toluenesulfonic acid monohydrate (0.68 g, 3.6 mmol) was addedin one portion. The solution was heated to 80° C. The mixture continuedto stir at 80° C. for a total of 1 h. The mixture was cooled to rt, andwithout concentrating the solution, the mixture was passed through aplug of silica/Celite eluting with ether and toluene. A black insolubleresidue remained clinging to the flask. Concentration of the eluent invacuo provided a dark tar. The material was purified by Biotage flashchromatography in three portions (50 g SNAP, 7-40% acetone/hexanes). Theappropriate fractions from each chromatography were combined andevaporated to give Δ23-24-dehydroascomycin as a white powder (4.0 g,61%).

Δ23-24-dehydroascomycin (0.54 g, 0.70 mmol) was dissolved in absoluteethanol (65 mL), and then methoxylamine hydrochloride (0.70 g, 8.4 mmol)was added followed by pyridine (0.56 mL, 7.0 mmol). The mixture wasstirred at 60° C. After 3.5 h, the reaction was cooled to rt and dilutedwith water. The ethanol was rotary evaporated. The aqueous residue wastreated with a saturated aqueous NaHCO₃ solution to adjust to pH 6, andthen extracted with EtOAc (3×50 mL). The combined organic phase waswashed with brine, dried over Na₂SO₄, and evaporated to give a whitesolid. Purification by Biotage FC (25 g SNAP, 7-60% Acetone/Hexane). Theappropriate fractions were combined and evaporated to give the desiredproduct as a white solid (0.29 g, 51%).

Preparation of C24-Deoxyascomycin

Ascomycin (6.8 g, 8.6 mmol) was dissolved in toluene (140 mL) and thenp-toluenesulfonic acid monohydrate (0.68 g, 3.6 mmol) was added in oneportion. The solution was heated to 80° C. The mixture continued to stirat 80° C. for a total of 1 h. The mixture was cooled to rt, and withoutconcentrating the solution, the mixture was passed through a plug ofsilica/Celite eluting with ether and toluene. A black insoluble residueremained clinging to the flask. Concentration of the eluent in vacuoprovided a dark tar. The material was purified by Biotage flashchromatography in three portions (50 g SNAP, 7-40% acetone/hexanes). Theappropriate fractions from each chromatography were combined andevaporated to give Δ23-24-dehydroascomycin as a white powder (4.0 g,61%).

The Δ23-24-dehydroascomycin (1.6 g, 2.0 mmol) was dissolved in methanol(25 mL) and added to a suspension of 10% palladium on carbon (0.12 g) inmethanol (25 mL). The flask was purged with nitrogen, then hydrogen. Aballoon with hydrogen was affixed to the flask with a needle through arubber septum. The mixture was stirred briskly for 18 min, beforecarefully filtering through a pad of Celite with MeOH (make sure to keepthe pad of Celite wet with MeOH). The solvent was evaporated to give agray foamy solid. Purification by Biotage flash chromatography (50 gSNAP, 7-60% acetone/hexane, collecting on threshold (30 mAu). Fractions4-5 were combined and evaporated to give C24-deoxyascomycin as a foamywhite solid (0.73 g, 46%).

Preparation of C24-deoxy-C22-hydroxy ascomycin.

Ascomycin (6.8 g, 8.6 mmol) was dissolved in toluene (140 mL) and thenp-toluenesulfonic acid monohydrate (0.68 g, 3.6 mmol) was added in oneportion. The solution was heated to 80° C. The mixture continued to stirat 80° C. for a total of 1 h. The mixture was cooled to rt, and withoutconcentrating the solution, the mixture was passed through a plug ofsilica/Celite eluting with ether and toluene. A black insoluble residueremained clinging to the flask. Concentration of the eluent in vacuoprovided a dark tar. The material was purified by Biotage flashchromatography in three portions (50 g SNAP, 7-40% acetone/hexanes). Theappropriate fractions from each chromatography were combined andevaporated to give Δ23-24-dehydroascomycin as a white powder (4.0 g,61%).

The Δ23-24-dehydroascomycin (1.6 g, 2.0 mmol) was dissolved in methanol(25 mL) and added to a suspension of 10% palladium on carbon (0.12 g) inmethanol (25 mL). The flask was purged with nitrogen, then hydrogen. Aballoon with hydrogen was affixed to the flask with a needle through arubber septum. The mixture was stirred briskly for 18 min, beforecarefully filtering through a pad of Celite with MeOH (make sure to keepthe pad of Celite wet with MeOH). The solvent was evaporated to give agray foamy solid. Purification by Biotage flash chromatography (50 gSNAP, 7-60% acetone/hexane, collecting on threshold (30 mAu). Fractions4-5 were combined and evaporated to give C24-deoxyascomycin as a foamywhite solid (0.73 g, 46%).

To a solution of C24-deoxyascomycin (0.33 g, 0.42 mmol) in THF (4 mL) at-70° C. was added K-Selectride (1.1 mL of 1.0 M soln in THF, 1.1 mmol)dropwise. The temperature remained at −70° C. to −40° C., and TLC at 4 hindicated no r×n. The clear/colorless soln was placed into the −20° C.freezer. It gradually turned yellow, then orange, over a 2 h period. Themixture was cautiously poured into a beaker of ice. Dilute HCl was addedto adjust to neutral pH (the solution became colorless). The mixture wasextracted with ethyl acetate (3×50 mL). The combined organic phase wassuccessively washed with water and brine, then dried over Na₂SO₄. Thevolatiles were evaporated to give a yellow oil. The mixture was purifiedby Biotage FC (25 g SNAP, 7-60% Acetone/Hexanes). The appropriatefractions were combined and evaporated to give the desired product as awhite solid (0.16 g, 48%).

Preparation of C22-oximes-C21-alkenes (other than allyl) fromTacrolimus.

Example Procedure using hydroxylamine and propene: FK506 (2.0 g, 2.5mmol) was dissolved in diethyl ether (40 mL), and the mixture wasde-gassed with nitrogen for 10 min.Dichloro[1,3-bis(2,6-isopropylphenyl)-2-imidazolidinylidene](benzylidene)(tricyclohexylphosphine)ruthenium(II) “Furstner catalyst” (30 mg) andCuI (20 mg) were then added, followed by liquid propene (5 mL, condensedfrom gas) and the mixture was stirred for 16 h at rt. Isolute Si-Thiolresin (Biotage) was added, and the mixture was stirred for 1 hr, thenallowed to stand. The supernatant was decanted, and the resin was washedwith ether (20 mL) and hexane (20 mL). The combined supernatants wereconcentrated in vacuo to an oily residue. This material was purified bypreparative HPLC to give the desired propenyl compound as a white solid.

The product from above (0.10 g, 0.12 mmol), hydroxylamine hydrochloride(0.017 g, 0.24 mmol), pyridine (1 mL), and ethanol (1 mL) were placed ina 4 mL vial and stirred overnight at rt. The ethanol was evaporated andthe residue was poured into 1M HCl (aq). The product was extracted withdichloromethane, and the solvent was evaporated to give a clear glassysolid. This material was purified by Biotage flash chromatography (10 gSNAP, 40% acetone/hexane). The appropriate fractions were collected,pooled and concentrated to give a glassy solid which was then dissolvedin acetonitrile/water and lyophilized to give the desired product (apair of isomers) as a white powder (10 mg, 10%).

Activity: Activity against C. neo, Candida, Candida w/FLu., Asp, andAsp/Caspo was determined using standardized in vitro susceptibilitytests; see, Clinical and Laboratory Standards Institute (CLSI) and theEuropean Committee for Antimicrobial Susceptibility Testing (EUCAST),and compounds 2-219 each demonstrate antifungal activity against one ormore of these fungi; exemplary, excerpted data are shown below.

Active C. Active Candida Active Asp Active Asp Compound neo w/FLu. alone(MEC) Caspo 2 yes no yes Yes 3 yes yes no yes 4 ug/mL 4 yes yes no yes 8ug/mL 5 yes yes yes yes (8 ug/mL) 1 ug/mL

Compounds most active against C. neo include #2-6, 8, 11, 14-18, 20,23-24, 26-28, 30-32, 35-44, 47, 50, 55, 58-80, 82, 86-91, 97-102, 116,118-120, 123, 127-128, 133-135, 138-150 and 152-161.

Compounds most active against Asp include #2, 5, 11, 15-18, 23, 24, 26,39, 52, 55, 79, 86, 87, 89, 97-101, 132-141, 144, 146-150, 152, 153,155-158, 160, 161, 163, 166, 174, 178, and 179-181.

IL2 IC₅₀ values were also determined, with compounds 2-219 demonstratingIC₅₀ in the subnanomolar (e.g. #42, 51, 61, 75-76, 103, 126, 129, 132,138, 140, 151, 163), nanomolar (e.g. #2, 3, 8, 18, 23-24, 31-32, 37,39-40, 44, 52, 60, 62-66, 68-69, 71-72, 77-79, 81-91, 96-102, 104-125,127, 130-131, 133-137, 139, 142-150, 152-154, 157-158, 160-162, 164-168,170, 172-175, 178-179, 184) and micromolar (e.g. #53, 55-57, 67, 70,73-74, 80, 112, 155, 177) ranges.

The invention encompasses all combinations of recited particular andpreferred embodiments. It is understood that the examples andembodiments described herein are for illustrative purposes only and thatvarious modifications or changes in light thereof will be suggested topersons skilled in the art and are to be included within the spirit andpurview of this application and scope of the appended claims. Allpublications, patents, and patent applications cited herein, includingcitations therein, are hereby incorporated by reference in theirentirety for all purposes.

What is claimed is:
 1. A compound selected from the group consisting ofcompounds 2-37, 39-91, 96-155, 157-158, 160-164, 167-190, 193, 200,202-213, 215, 217, 218, and 219 as recited in Table
 1. 2. Apharmaceutical composition comprising a formulation according to claim 1in effective unit dosage.